Protocol for generating postganglionic sympathetic neurons using human pluripotent stem cells for electrophysiological and functional assessments

Summary

Assessing the development and function of the sympathetic nervous system in diseases on a large scale is challenging. Here, we present a protocol to generate human pluripotent stem cell (hPSC)-derived postganglionic sympathetic neurons (symNs) differentiated via neural crest cells (NCCs), which can be cryopreserved. We describe steps for hPSC replating, NCC replating and cryobanking, and symN differentiation. We then demonstrate the functionality of the hPSC-derived symNs, focusing on electrophysiological activity, calcium flux, and norepinephrine dynamics. For complete details on the use and execution of this protocol, please refer to Wu et al.1,2. Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.

Authors Wu HF, Art J, Saini T, Zeltner N
Journal STAR protocols
Publication Date 2024 Jun 21;5(2):102970
PubMed 38517897
PubMed Central PMC10966798
DOI 10.1016/j.xpro.2024.102970

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